INSTRUMENTATION FOR PROTEOMICS

在这项提案中，我们请求支持购买一个仪器系统，以半自动处理在弗吉尼亚大学生物分子研究设施进行的蛋白质组项目的样本。具体地说，我们需要一个凝胶成像系统和相关的软件，凝胶斑点拾取器，凝胶斑点消化器和消化仪。这些仪器产生的样品将在我们现有的PE Voyager DE Pro MALDI-TOF仪器上进行自动质量测绘，或者如有必要，使用该设施的Finnigan LCQ离子陷阱LC/MS/MS质谱仪进行从头测序。该计划的总体目标是为蛋白质组相关项目的研究人员提供高通量、低成本的蛋白质鉴定。此外，这项技术将与该设施中的微阵列/基因芯片计划对接。这些仪器将被安置在弗吉尼亚大学生物分子研究设施中，并将支持七名由NIH资助的研究人员的蛋白质组研究项目。对于这些项目，我们预计这些仪器将使用大约80%的可用时间。剩余的时间将用于新的蛋白质组相关项目的开发。该大学没有其他的高通量或其他的质谱学蛋白质鉴定服务。生物分子研究机构主任杰伊·福克斯博士将负责仪器的全面管理。尼古拉斯·谢尔曼博士将负责蛋白质组项目的质谱学问题，约翰·香农博士将负责蛋白质组项目的蛋白质化学方面。两人都是生物分子研究机构的工作人员。这三位科学家将在蛋白质组学项目的规划、执行和数据分析方面与调查人员进行咨询。在过去的七年里，该设施提供了质谱学蛋白质鉴定和从头序列分析，预计样品分析的下游端不会出现重大困难。将实行按服务收费，以帮助支付工作人员工资和用品费用。生物分子研究设施的监督由医学院院长任命的一个常设委员会提供。监督委员会直接向福克斯博士提供指导和专门知识，以管理该设施，并补充设施工作人员的科学专长。该提案的主要调查人员之一凯文·哈森博士是该委员会的成员之一。弗吉尼亚大学生物分子研究设施在过去20年里一直在弗吉尼亚大学健康科学中心的持续财政支持下运行。健康科学中心的管理层致力于该设施的长期运营。这一支持，以及监督委员会和设施工作人员的支持，将有助于确保充分、适当地使用仪器，以确保为弗吉尼亚大学NIH资助的调查人员所需的仪器的财政健全和科学上的成功运作。

In this proposal, we are requesting support for the purchase of an instrument system to semi-automate sample processing for proteomic projects performed in the University of Virginia Biomolecular Research Facility for University investigators. Specifically, we are requesting a gel imaging system and associated software, gel spot picker, gel spot digester and digest spotter. Samples generated by these instruments will be subjected to automated mass mapping on our existing PE Voyager DE Pro MALDI-TOF instrument or if necessary, de novo sequencing with the facility's Finnigan LCQ ion-trap LC/MS/MS mass spectrometer. The overall goal of this program is to provide high throughput, low cost protein identification for investigators with proteomic related projects. Furthermore, this technology will be interfaced with the Microarray/GeneChip Program in the facility. The instruments will be housed in the University of Virginia Biomolecular Research Facility and will support the research proteomic projects of seven NIH-funded investigators. For these projects, we anticipate that the instruments will be in use approximately 80% of the available time. The remaining time will be available for new proteomic-related project development. There is no other mass spectrometric protein identification service, high throughput or otherwise, available at the University. Dr. Jay Fox, Director of the Biomolecular Research Facility, will be responsible for the overall management of the instrumentation. Dr. Nicholas Sherman will be responsible for mass spectrometry issues of proteomics projects and Dr. John Shannon will be responsible for the protein chemistry aspects of the proteomic projects. Both are staff members in the Biomolecular Research Facility. All three scientists will consult with investigators in the planning, execution and data analysis of proteomics projects. For the past seven years, the facility has offered mass spectrometric protein identification and de novo sequence analysis and no major difficulties are expected on the downstream end of sample analysis. A fee-for-service charge will be instituted to help defray the costs of staff salaries and supplies. Oversight of the Biomolecular Research Facility is provided by a standing committee appointed by the Dean of the Medical School. The Oversight Committee provides guidance and expertise directly to Dr. Fox for the management of the facility as well as complementing the scientific expertise of facility staff. One of the principal investigators in this proposal, Dr. Kevin Hazen, is a member of the committee. The University of Virginia Biomolecular Research Facility has been in operation for the past 20 years with continual fiscal support from the University of Virginia Health Sciences Center. The administration of the Health Sciences Center is committed to the long-term operation of the facility. This support, along with that of the Oversight Committee and facility staff, will serve to ensure adequate, appropriate instrument use for the fiscally sound and scientifically successful operation of instrumentation requested for the benefit of NIH-funded investigators at the University of Virginia..